The present invention is directed to an apparatus and method for collecting a uniform monolayer of particulate matter. In particular, the present invention is directed to an apparatus and manual or semi-automatic method for collecting a uniform monolayer of cells from biological fluids and preparing the monolayer of cells for use in cytological protocols.
In a wide variety of technologies, the ability and/or facility for separating matter, typically particulate matter, from a fluid is a critical component in the ability to test for the presence of substances in the fluid. Too often, interference associated with sample preparation obscures the target cells to such a degree that the process is not sufficiently reliable, or too costly.
Such a scenario applies to many other fields which involve detection and/or diagnosis, including environmental testing, radiation research, cancer screening, cytological examination, microbiological testing, and hazardous waste contamination, to name just a few.
In all of these endeavors, limiting factors in the sample preparation protocol include adequately separating particulate matter from its fluid carrier (e.g., physiological fluid, biological fluid and environmental fluid), and easily and efficiently collecting and concentrating the particulate matter in a form readily accessible for microscopic examination.
In the case of cytological examination, a sample of cells is obtained from a patient. Typically, this is done by scraping or swabbing an area, as in the case of cervical samples, or by collecting body fluids, such as those obtained from the chest cavity, bladder, or spinal canal, or by fine needle aspiration. In a conventional manual cytological examination, particulate matter including cells and debris in the fluid are transferred onto a glass slide by smearing and subsequently air-dried. Smearing results in non-uniform densities and uneven distributions of cells and debris that often obscure the target cells. Air drying causes cell distortion and further impedes accurate examination.
It has been found that prompt processing of urine to obtain fresh ensures the accuracy of quantitative culture results, urinalysis and microscopy. Fresh cells tend to stick to a glass slide much better than cells from preserved urine, allowing for smoother cell spread onto the glass body. Delays in processing, negligent care in either inpatient or outpatient settings and lack of refrigeration may lead to non-optimal slide preparation. One known solution to the delay problem is the use of chemical preservatives with the urine. The presence of liquid preservatives, however, in the urine specimen raises the specific gravity of the specimen to unmeasurable levels and may limit the potential usefulness of the urine for various types of traditional quantitative analysis, such as slide microscopy.
Diagnostic microbiology and/or cytology, particularly in the area of clinical pathology, bases diagnoses on a microscopic examination of cells and other microscopic analyses. The accuracy of the diagnosis and the preparation of optimally interpretable specimens typically depends upon adequate sample preparation. New methodologies such as immunocytochemistry and image analysis require preparations that are reproducible, fast, biohazard-free and inexpensive. Conventional cell preparation techniques fail to adequately address the issues of non-uniform cell densities, uneven cell distribution and air drying artifacts.
Conventionally, body fluid samples are collected for cytological examinations using containers that contain a preservative solution for preserving the cytology specimen during shipment from the collection site to the cytology laboratory. Furthermore, cytology specimens collected from the body cavities using a swab, smear, flush or brush are also preserved in containers with fixatives (e.g., alcohol or acetone fixatives) prior to transferring cells onto the slide or membrane for staining or examination.
It is desirable to provide a urine or other biological fluid specimen container that would allow liquid biological specimens to be tested without removing the lid of the urine or biological fluid container. However, none of the prior art solves the problems of transferring cells in a monolayer to a slide for examination without submerging portions of the device in the sample (and increasing the risk of contamination), consistently and repeatedly forming a high quality monolayer on the microscope slide, and processing the sample so that the fluid from which the cells were taken is preserved.
A number of methods, apparatuses, and structures for dispersing cells in the fluid are known. For example, U.S. Pat. No. 5,143,627 opens the sample container, inserts a dispersing element into the liquid suspension, and rotates the dispersing element for several minutes. In another example, the so-called xe2x80x9cSaccomanno methodxe2x80x9d is used to process sputum, a process that is time consuming and involves a large number of processing steps.
In contrast to the conventional techniques, the solid matter preparation techniques of the present invention address the issues of non-uniform matter densities, uneven matter distribution, and sample loss and contamination due to the number of steps involved in the sample preparation. Thus, preparations according to the present invention result in an even distribution of solids that have superior morphology, improved visualization, and are readily positioned and available for light absorbance analysis without the need to further manipulate or prepare the sample.
The present invention relates to an apparatus and method for collecting matter for detection, analysis, quantification, and/or visualization. The devices and methods of the present invention are particularly suitable for separating particulate matter from biological, physiological, and environmental fluids and presenting the particulate matter in an improved manner for cytological examination.
A preferred embodiment of the present invention relates to an apparatus and method for collecting a uniform layer of cells from urine or other biological fluid specimen in a cytology collection apparatus or assay module, and for transferring the uniform layer of particulate matter to a slide.
The devices and methods of the present invention may be configured into a hand-held manual system or structure, or a partially automated system or structure.
Such an apparatus according to the present invention overcomes the problems associated with conventional equipment for collecting cells and other particles for cytology by providing a mechanism of relatively simple structure and operation that separates particles from a liquid solution, collects an approximately known quantity of the cells in a monolayer, and transfers the collected cells to a microscope slide. In some embodiments of the present invention, no element of the apparatus is placed in the liquid sample, thus preventing unnecessary contamination of the sample. Moreover, in some embodiments of the present invention, the container holding the sample is not opened in the course of collecting and transferring the cells, thus eliminating the possibility of sample contamination during testing.
In all embodiments of the present invention, a monolayer of the particulate matter, e.g., cells, in the sample is collected on a filter by passing two branches of a fluid flow through and around the filter. Such a filter is known from U.S. Pat. Nos. 5,301,685 and 5,471,994, which are incorporated by reference.
The patient or medical person handling the collection may seal a separate container. The collection of the cells according to the present invention allows a uniform cell slide to be obtained without contamination of the cells by preservatives, workers or outside materials. The transfer from collection container to the cytology collection apparatus may be carried out without pouring or pipetting the collected specimen.
The present invention is directed to a cell collection and distribution apparatus that can be disassembled to allow face to face transfer of cells from the device to a slide for microscope examination. The present invention provides an improved apparatus and method for collecting a monolayer of cells that can be transferred to a microscope slide. The effectiveness of transferring the monolayer cells from the filter to a microscope slide has proven to be very high without differential cell loss. Microscopic examination shows that the cell distribution is the same on the slide as on the filter.
The devices of the present invention obviate the need for a trained technician to properly prepare a sample substrate. Thus, time, expense, and expertise are eliminated or reduced as critical factors in sample preparation protocols.
The devices and methods of the present invention also provide advantages in sample preparation because they are suitable for use with fresh, untreated cells, unmodified cells, and are particularly designed to provide a thin, uniform layer of solid matter (up to approximately 40 microns or more). This invention is particularly useful for collecting cells for a Pap smear.
The apparatuses and methods of the present invention have many advantages for conventional microbiology and hematology. The collected cells are in a pre-determined area that is easily accessible to a radiant light source and to a wavelength absorbance meter. Because cells are concentrated in a single layer, they are almost always in one focal plane, thus eliminating or reducing interference by other particles and virtually eliminating technician time and expertise in establishing a proper reading. The minimal matter overlap achieved by the present invention ensures that all matter can be easily examined with little chance for critical solids to be obscured by clumps of overlapping solids or debris. Certain embodiments of the apparatuses of the present invention may be used in combination with other automated devices to detect and analyze any solid matter in a given population. They also permit a detailed analysis of the chemical composition of the matter.
The present invention also includes an improved apparatus and method for processing a fluid containing particulate matter. The apparatus and method include dispersing particulate matter in the sample, preferably by rotating the sample container around a fixed agitator or by rotating the agitator inside a fixed sample container. The present invention agitates the sample within the container to ensure break-up of large particulate matter, e.g., mucoid bodies in the case of sputum samples, and the even distribution of cells throughout the fluid. Agitation may occur as the result of relative motion between components of the sample container, non-uniform motion of the sample container, and/or inertial reaction forces applied to the sample by the container.
According to a preferred embodiment of the present invention structures and means are provided for rotating an agitator in relation to the container and/or the sample in the container. As described in more detail below, a preferred embodiment according to the present invention may include a cover within a cover, wherein the agitator is fixed to a freely rotatable outer cover and an inner cover is secured with respect to a stationary sample container. Such relative motion moves the agitator in relation to the sample, and disperses particulate matter in the fluid.
Further, providing a container cover that has a portion that is rotatable permits particulate matter stirring or dispersion without inserting a stirring mechanism into the sample, thus eliminating a source of contamination that plaques devices that are presently commercially available. In preferred embodiments of the present invention, the covering on the sample container may include a hollow tube, with or without a rotatable dispersing element, for withdrawing the sample from the container.
In a preferred embodiment of the invention, the cover comprises a first portion that fixedly engages the container and a second portion that may be rotatable in relation to the container. As used herein, rotatable in relation to the container refers to the relative movement of the first portion and the second portion; the first portion may be fixed and the second portion moveable, or the first portion may be moveable and the second portion fixed. In a most preferred embodiment, the second or inner portion of the cover is stationary and the first or outer portion is rotatable. In a preferred embodiment of the invention, the agitator is engaged by or fixed to the second portion of the cover.
An apparatus according to a preferred embodiment of the present invention may be configured to support, engage, and rotate a portion of a collection container so that the sample is mixed according to the present invention. An exemplary collection container includes a container or cup suitable for collecting and holding a specimen sample, a cap having a first position that is not rotatable in relation to the container and a second position that is rotatable in relation to the container, and an agitator engaged by or fixed to a portion of the cover and extending into the container. As used herein, configured to support, engage, and rotate refers to various configurations that may be adapted to perform the specific function. For example, an apparatus according to the invention may include a container support for positioning at least one sample container and rotating the container per se, and a sleeve or clamp for engaging and fixing a portion of the cap that communicates with an agitating element. Alternatively, the support may hold the container in a fixed position and a pulley, sleeve, or clamp may engage and rotate the portion of the cap that is fixed with respect to an agitator. In a preferred embodiment of the invention, a sleeve engages an inner portion of the cap, and holds the inner portion of the cap in a stationary position in relation to an outer portion of the cap.
Configurations or structures that engage a portion of the cap or the container typically include any member that positions, fixes and/or moves either that portion of the cap or the container. Exemplary members include, but are not limited to, a sleeve, one or more belts, one or more pulleys, one or more resilient bands, and the like.
The present invention is also a device for processing a fluid into one or more components, typically by removing particulate matter from the fluid. The present invention is directed to apparatuses and methods for collecting fluids, such as biological, physiological, or environmental fluids, and removing particulate matter from the fluid, without centrifugation, and diagnosing and testing the matter. In a preferred embodiment of the invention, particulate matter is collected on a collection site. In a most preferred embodiment of the invention, the particulate matter is collected in a monolayer and in a pre-determined spatial arrangement.
While a cytology collection apparatus according to the invention can be used for any biological fluid, it is particularly useful for preparing testing samples from urine and its associated cells for Pap smears. It is intended that the type of matter being processed should not limit the invention. In a most preferred embodiment of the invention, the fluid is urine and particulate matter is a cell. The particulate matter processing apparatus of the present invention also permits isolation and collection of fresh cells and/or microorganisms from biological fluids to perform DNA probe and chromosomal analysis once the proper buffer hemolyzes the cells.
In the case of cervical examinations, a scraping of the cervix is taken with a longhandled brush or broom. The handle is then shortened, such as by breaking or telescopic movement, and the brush is inserted into a specimen container. Conventionally, the container must be opened to remove the brush at the time of testing. Such a process increases the likelihood of contamination because the cover of the sample container must be opened, the brush typically retains cells if the testing is not performed soon after cell collection, and the operator must come into contact with the sample.
According to a preferred embodiment of the present invention, these problems are avoided by providing a system in which the brush not only remains in the collection container, but can also be used to disperse the collected cells during agitation. Further, the apparatus of the present invention is a closed system; once the apparatus is closed, it does not need to be opened in order to process any cells collected on the brush.
Further, providing a container cover that has a portion that is rotatable permits particulate matter stirring or dispersion without inserting a stirring mechanism into the sample, thus eliminating a source of contamination that plaques devices that are presently commercially available.
The present invention is also directed to a cytology collection and testing kit containing a cytology collection apparatus, replacement filters, replacement disposables, and/or other components, ingredients of a fixative composition as described below. The cytology collection kit may also include replacement filters, replacement disposables, and/or other components, ingredients or solutions typically used during cytological examinations. The kit might also include washing, fixative, and/or buffer solutions. A cervical kit may include a brush or broom, and a fluid suitable for storing the used brush until particulate matter on the brush can be processed through the filter assembly.
According to another aspect of the present invention, the matter collection apparatus may also include additional modules, removable or integrated, for treating the fluid. For example, the fluid may be treated with a matter collection module, in combination with a debris removal module, a chromatography module, and assay module, or combinations of these and other devices. These and other modules or treatment protocols provide features that may be desirable to incorporate into a sample preparation apparatus according to the invention.
The devices and methods of the present invention have many advantages for conventional cytology. The cells are in a pre-determined area allowing for significant timesaving when screening the slide. Such problems as cells lying outside the coverslip or on the frosted end are eliminated. Because cells are lying in a single layer, they are almost always in a one focal plane when using a 10xc3x97objectivexe2x80x94the objective most often used for the lower power screening of a slide. Even with a 40xc3x97objective, most cells are in focus. This eliminates frequent refocusing and saves time.
The objects of the present invention are achieved by an apparatus for simultaneously processing a positive whole number of samples wherein each of the samples includes a fluid containing a respective particulate matter. The apparatus comprises a number of containers corresponding to the number of samples, each of the containers being adapted for holding a respective sample; a number of pumps corresponding to the number of samples, each of the pumps being adapted for communicating a respective fluid with a respective container; a number of filters corresponding to the number of samples, each of the filters being interposed between a respective pump and its respective container and being adapted for collecting a respective particulate matter; a first engagement supporting each of the containers, the first engagement having a number of first receivers corresponding to at least the number of samples, each of the containers being contiguously engaged by a respective first receiver; and a second engagement retaining the pumps, the second engagement having a number of second receivers corresponding to the number of first receivers, each of the pumps being contiguously engaged by a respective second receiver. Relative movement between respective ones of the first and second receivers disperses a respective particulate matter in its respective fluid.
The objects of the present invention are also achieved by a method for simultaneously processing a number of samples wherein each of the samples are held in a respective container and include a respective fluid containing a respective particulate matter. The method comprises closing each of the containers with a respective pump, each pump including a respective filter being interposed between its respective container and its respective pump and being adapted for collecting its respective particulate matter; supporting each of the containers on a first engagement, the first engagement having a number of first receivers corresponding to at least the number of samples, each of the containers being contiguously engaged by a respective first receiver; retaining the pumps on a second engagement, the second engagement having a number of second receivers corresponding to the number of first receivers, each of the pumps being contiguously engaged by a respective second receiver; and moving respective ones of the first receivers relative to respective ones of the second receivers for dispersing its respective particulate matter in its respective fluid.
The accompanying drawings show illustrative embodiments of the invention from which these and other of the objectives, novel features and advantages will be readily apparent.